LOCUS       V022656                 7430 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022656
VERSION     V022656
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2920
                     /label="MMP2(NM_004530)"
                     /note="MMP2(NM_004530)"
                     /gene="MMP2"
     CDS             2927..2992
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3030..3254
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3300..3728
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3742..4071
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4138..4932
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5106..5227
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5276..5292)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5300..5316
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5324..5354)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5369..5390
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5678..6263)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6434..7294)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7295..7399)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggaggcgc taatggcccg
      961 gggcgcgctc acgggtcccc tgagggcgct ctgtctcctg ggctgcctgc tgagccacgc
     1021 cgccgccgcg ccgtcgccca tcatcaagtt ccccggcgat gtcgccccca aaacggacaa
     1081 agagttggca gtgcaatacc tgaacacctt ctatggctgc cccaaggaga gctgcaacct
     1141 gtttgtgctg aaggacacac taaagaagat gcagaagttc tttggactgc cccagacagg
     1201 tgatcttgac cagaatacca tcgagaccat gcggaagcca cgctgcggca acccagatgt
     1261 ggccaactac aacttcttcc ctcgcaagcc caagtgggac aagaaccaga tcacatacag
     1321 gatcattggc tacacacctg atctggaccc agagacagtg gatgatgcct ttgctcgtgc
     1381 cttccaagtc tggagcgatg tgaccccact gcggttttct cgaatccatg atggagaggc
     1441 agacatcatg atcaactttg gccgctggga gcatggcgat ggatacccct ttgacggtaa
     1501 ggacggactc ctggctcatg ccttcgcccc aggcactggt gttgggggag actcccattt
     1561 tgatgacgat gagctatgga ccttgggaga aggccaagtg gtccgtgtga agtatgggaa
     1621 cgccgatggg gagtactgca agttcccctt cttgttcaat ggcaaggagt acaacagctg
     1681 cactgatacc ggccgcagcg atggcttcct ctggtgctcc accacctaca actttgagaa
     1741 ggatggcaag tacggcttct gtccccatga agccctgttc accatgggcg gcaacgctga
     1801 aggacagccc tgcaagtttc cattccgctt ccagggcaca tcctatgaca gctgcaccac
     1861 tgagggccgc acggatggct accgctggtg cggcaccact gaggactacg accgcgacaa
     1921 gaagtatggc ttctgccctg agaccgccat gtccactgtt ggtgggaact cagaaggtgc
     1981 cccctgtgtc ttccccttca ctttcctggg caacaaatat gagagctgca ccagcgccgg
     2041 ccgcagtgac ggaaagatgt ggtgtgcgac cacagccaac tacgatgatg accgcaagtg
     2101 gggcttctgc cctgaccaag ggtacagcct gttcctcgtg gcagcccacg agtttggcca
     2161 cgccatgggg ctggagcact cccaagaccc tggggccctg atggcaccca tttacaccta
     2221 caccaagaac ttccgtctgt cccaggatga catcaagggc attcaggagc tctatggggc
     2281 ctctcctgac attgaccttg gcaccggccc cacccccacg ctgggccctg tcactcctga
     2341 gatctgcaaa caggacattg tatttgatgg catcgctcag atccgtggtg agatcttctt
     2401 cttcaaggac cggttcattt ggcggactgt gacgccacgt gacaagccca tggggcccct
     2461 gctggtggcc acattctggc ctgagctccc ggaaaagatt gatgcggtat acgaggcccc
     2521 acaggaggag aaggctgtgt tctttgcagg gaatgaatac tggatctact cagccagcac
     2581 cctggagcga gggtacccca agccactgac cagcctggga ctgccccctg atgtccagcg
     2641 agtggatgcc gcctttaact ggagcaaaaa caagaagaca tacatctttg ctggagacaa
     2701 attctggaga tacaatgagg tgaagaagaa aatggatcct ggcttcccca agctcatcgc
     2761 agatgcctgg aatgccatcc ccgataacct ggatgccgtc gtggacctgc agggcggcgg
     2821 tcacagctac ttcttcaagg gtgcctatta cctgaagctg gagaaccaaa gtctgaagag
     2881 cgtgaagttt ggaagcatca aatccgactg gctaggctgc ctcgaggact acaaagacca
     2941 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     3001 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3061 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3121 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3181 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3241 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3301 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3361 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3421 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3481 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3541 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3601 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3661 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3721 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3841 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3901 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3961 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4021 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4081 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4141 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4201 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4261 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4321 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4381 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4441 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4501 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4561 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4621 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4681 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4741 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4801 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4861 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4921 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4981 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5041 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5101 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5161 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5221 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5341 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5461 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5521 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5581 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5641 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5701 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5761 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5821 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5881 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5941 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6001 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6061 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6121 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6181 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     6241 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6301 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6361 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6421 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6481 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6541 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6601 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6661 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6721 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6781 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6841 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6901 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6961 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7021 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7081 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7141 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7201 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7261 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7321 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7381 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
